Multiple endocrine neoplasia (MEN) is dominantly inherited tumor syndromes characterized by multiple endocrine hyperfunctions due to hyperplastic or adenomatous as well as carcinomatous abnormalities. The syndromes are classified into MEN 1, 2A and 2B according with the complex of abnormalities. Hyperparathyroidism is the most common manifestation of MEN 1 and is also associated with MEN 2A. Recent genetic studies revealed that alleleic deletions on chromosome 11q13 are involved in the monoclonal development of endocrine cell in MEN 1 and germ-line mutations of RET proto-oncogene are associated with MEN 2A and 2B. This paper reviews clinical characteristics of MEN 1, 2A and 2B in Japan and presents the respective case reports including the gene analysis.

Genomic imprinting is defined as a differential expression of genes depending on their parental origin. IGF2 gene on chromosome 11p, which is maternally imprinted in normal human development, has been demonstrated to be activated by loss of imprinting (LOI) during tumorigenesis of Wilms tumor or other kidney neoplasms in childhood. Although the molecular mechanism of genomic imprinting is not clarified yet, it is certain that disruption of the imprinting mechanism has a causative role in some human cancers.

Loss of heterozygosity on chromosome 16q region is shown to occur frequently in breast cancers regardless of difference in phenotype or extent of tumor spread, and inactivation of tumor-suppressor genes located on the region is considered to be involved in the acquisition of malignant phenotype of mammary glandular epithelial cells. Intracystic papillary carcinoma of the breast is diagnostically important entity because of the difficulty of differential diagnosis from intraductal papilloma preoperatively. Indeed, intracystic papillary carcinoma is usually of low-grade histologic atypia (Grade 1), and its well differentiated nature is biochemically supported by high estrogen-receptor and progesterone-receptor value in their tissue extract. Among intracystic papillary tumors of the breast, LOH on chromosome 16q was detected in 71% of intracystic papillary carcinoma, whereas it was absent in intraductal papilloma by both of Southern blot analysis and microsatellite polymorphism analysis mediated by polymerase chain reaction. Therefore, LOH on chromosome 16q was suggested to be helpful for differential diagnosis of intracystic papillary tumors and to be applicable to the materials of fine needle aspiration performed preoperatively.

To clarify characteristics and development of stump cancers of the stomach, we studied 10 cases (12 lesions) of them with mucin-histochemical and immunohistochemical techniques and gene analysis using polymerase chain reaction. Sixty-seven % of the cancers were mostly composed of gastric-type cells and 67% also showed abnormal accumulation of p53 protein in their nuclei. There were scattered cells with abnormal accumulation of p53 protein in cystically dilated glands that were often found to be surrounding cancers. Immunohistochemistry of proliferating cell nuclear antigen also demonstrated proliferating activity of these cystic glands. It is suggested that the cystically dilatated gland is precancerous lesion of stump cancers of the stomach. The gene analysis showed less occurrence of K-ras abnormality, and the mutation of APC gene is suggested to be infrequent.

We studied the phenotypic expression, DNA ploidy patterns and other clinico-pathological features of early gastric cancers accompanied by lymph node metastasis. Of 241 cases studied, 18 (7.5%) were lymph node-positive. Histologically, 4 were tub1, 7 tub2, 4 sig and 3 por. Seven of the 18 were differentiated type cancers with accompanying undifferentiated type cancer cells in a part of the cancer tissue. Mucin histochemistry showed that 15 of the 18 cancers were composed exclusively of gastric type cells, while the other 3 were composed of intestinal type cells. The incidence of gastric type cells in node-positive cancers was significantly higher (p < 0.05) than that in node-negative cancers. All node-positive mucosal cancers were composed mainly of gastric type cells. These findings indicate that there are many cancers which are originally gastric in type, and that gastric type cancers readily develop lymph node metastasis. On the other hand, a cytofluorometric study showed that 10 tumors were diploid and 8 were aneuploid. There was no statistically significant association between DNA ploidy patterns and the occurrence of lymph node metastasis.

The value of tumor angiogenesis, EGFR and c-erbB-2 oncoprotein, a long with p 53 protein expression for predicting relapse-free survival was investigated in 110 node-negative breast cancer patients. The grade of neovascularization was assessed by the microvessel density which was obtained by an immunocytochemical staining by factor VIII-related antigen. EGFR, c-erbB-2 oncoprotein and p 53 oncoprotein were also determined by immunocytochemical assay. Univariate analysis showed no statistical significance of EGFR, c-erbB-2 and p53 status as a prognostic indicator. However, the microvessel density was a significant predictor of relapse-free survival. Patients with over 100 counts of factor VIII-RA positive cells per mm2 field in the most active areas of neovascularization showed significantly poorer prognosis compared to those with less than 100 counts (p < 0.005). Multivariate analysis demonstrated that microvessel density was an independent prognostic indicator in node-negative breast cancer patients (p < 0.0005). It was suggested that microvessel density might be of use in selecting the high-risk group in node-negative breast cancer patients needing adjuvant therapies.

In the previous studies, epidermal growth factor (EGF) was found to elevate glycolipid sulfotransferase activity in a human renal cell carcinoma cell line, SMKT-R3. To elucidate whether Ras is involved in the signal transduction pathway from EGF to the expression of the sulfotransferase, effects of EGF and a tyrosine kinase inhibitor on the sulfotransferase activity were investigated in renal cancer cells stably expressing activated Ras. SMKT-R3 cells were transfected with a plasmid carrying v-H-ras (pv-H-ras). As a control, SMKT-R3 cells transfected with a mutant v-H-ras, in which glycine at position 15 was replaced by valine (pG15V), was used. The expression of v-H-ras in transfected cells was examined by RT-PCR analysis. Two clones transfected with pv-H-ras, named A1 and A6, and two clones transfected with pG15 V, termed B1 and B4, were found to express v-H-ras and G15V genes, respectively, and employed in the following experiments. Though EGF elevated the activity of glycolipid sulfotransferase in B1, B4 and SMKT-R3 cells, it did not change the activity levels in A1 and A6 cells. This result suggested that since the signal from Ras was saturated in the cells expressing activated Ras, the cells did not respond to the stimulation by EGF. To know the association between tyrosine kinase of EGF receptor and Ras, the effect of Genistein, a specific inhibitor of tyrosine kinase, on the sulfotransferase activity was examined in these cell lines. Genistein reduced the activity of glycolipid sulfotransferase in B1, B4, and SMKT-R3 cells, whereas it hardly affected the sulfotransferase activity in A1 and A6 cells. This result indicated that the signal from tyrosine kinase was dissociated from the regulation of the sulfotransferase activity in the cells expressing activated Ras. These observations suggest that Ras is involved in the signal transduction from EGF to the expression of glycolipid sulfotransferase activity in SMKT-R3 cells and acts in the downstream of tyrosine kinase of EGF receptor.

Cytogenetic studies were performed on 25 patients with multiple myeloma. Five of 25 patients (20%) had chromosomally abnormal clones. The most common anomalous chromosomes were #1, #10, #12, #3, and #14. Chromosomal abnormalities of t (1 ; 10) (q42 ; q26) and t (1 ; 19) (q11 ; p13.3) were observed consistently in a Ig D myeloma patient. Patients with abnormal karyotypes were considered to be in a clinically aggressive phase because of high percentage of immature plasma cell in bone marrow and to have a poor prognosis. Current status of the chromosome findings in multiple myeloma are reviewed in this article.

Multiple myeloma (MM) is a malignant proliferation of bone marrow plasma cells. Molecular analyses of the involvement of oncogenes (c-myc, and N- and K-ras genes) and suppressor gene (p53) in pathogenesis of MM have been recently carried out. Relatively high incidence of elevated expression of c-myc mRNA have been found, although the gene rearrangements are very rare. Mutations of N- and K-ras genes have been found in about 1/3 of patients with MM. Point mutations of p53 gene were detected in about 10-20% of patients. The mutations were found in terminal or leukemic phase of the disease. These indicate that oncogenes and suppressor genes are involved in the development and progression of MM.

A moderately-differentiated endometrial adenocarcinoma cell line(EI) was established from a surgical specimens obtained from a 55-year-old woman with endometrial carcinoma. This cell line could be transplanted to nude mice, where the cells showed the same histological type as the primary tumor. The doubling time of the cell line was 50.5 hours; the saturation density was 7.5 x 104 cells/cm2; the plating efficiency was 46%. This cell line was determined to produce TPA, but not other tumor markers, such as CA125 or CEA. Neither estrogen receptor, nor progesterone receptor was detected from the culture cell or the primary tumor. Chromosome analysis revealed that cells examined were all 46,XX, + 8,t(14q14q), and only cells with this karyotype were thought to be able to grow. From these results, it was suggested that a gene on No. 8 chromosome would be involved in the carcinogenesis of endometrial adenocarcinoma. Thus this cell line was thought to be useful for the clarification of gene conversion during the process of development of endometrial adenocarcinoma.

A childhood embryonal rhabdomyosarcoma cell line, designated as TS-RM-1, was established from transplanted tumor in nude mouse. TS-RM-1 cells were small, spindle to polygonal shaped and cytoplasm was rich in glycogen. Estimated population doubling time was 31 hours and the distribution of chromosome number was in the range of 88 to 98. The karyotype of TS-RM-1 cells revealed nonrandom structural chromosome alterations, including der(3)t(1;3)(q12;p12-14),16q-,17q+ and 21q+. In immuno-cytohistochemical study, both TS-RM-1 cells and the primary tumor were positive for desmin and vimentin. TS-RM-1 cell line may be useful for studying the association between embryonal rhabdomyosarcoma and a specific alteration in chromosome 3.

The incidence of the point mutation of K-ras oncogene at codon 12 in surgical or autopsy specimens of pancreatic carcinoma (PC) is reportedly extremely high ranging from 75 to over 90%, and the K-ras mutation found in PC is almost exclusively present in codon 12. The GGT to GAT transition of the 12th codon is observed in more than 50% of PC of Japanese patients, although the incidences of transition of GGT to CGT, GTT, and GAT are essentially equal in PC of European countries. With this point in mind, the authors attempted to detect K-ras mutations in DNA obtained from pancreatic juice (PJ) collected endoscopically. K-ras mutations at codon 12 were found in 55% of 20 PC patients by PCR and ASO probe hybridization method, and in 80% of 25 PC patients by PCR-RFLP method. On the other hand, K-ras mutations were negative in PJ from chronic pancreatitis with one exception by PCR-RFLP method. Other authors also reported almost the same results. These results suggest that analysis of K-ras oncogene in PJ can be useful for qualitative diagnosis of PC, and would be expected as a novel tool for early detection of PC.

The Fas antigen is a cell surface protein that can mediate apoptosis, and plays a major role in functional maturity in clonal deletion of autoreactive T cells in the thymus. Recently a cDNA encoding the human Fas antigen was isolated. It was suggested that a signal-transducing domain, along with inhibitory one, was presented in the cytoplasmic domain of the Fas antigen. We examined whether any abnormality in Fas antigen gene may breakdown the immunotolerance in patients with myasthenia gravis (MG). We used single-strand conformational polymorphism (SSCP) analysis, generally used to screen for unknown mutation, to examine the cytoplasmic cDNA of Fas antigen obtained by reverse transcription (rt)-PCR from mRNA of MG thymuses. Furthermore, we studied in situ expression of Fas antigen mRNA in thymuses from MG patients. Fragments from all the 12 (3 control subjects, and 9 MG patients) thymuses produced by rt-PCR showed equally stained two bands of the two single strands. In the thymuses from both controls and MG patients, the Fas antigen mRNA was mainly expressed in small thymic cells. These cells were located near clustered cells with relatively large cytoplasms in the cortex and sometimes surrounded them, but were also founded in clusters in the follicules. In situ expressions of Fas antigen mRNA were more remarkable in MG thymuses than in control subjects, and in hyperplastic thymuses than in normoplastic ones. These results suggest that mRNA for the cytoplasmic domain of the Fas antigen have no mutation, and its expression is not reduced in thymuses from patients with MG.

We report a case of acute myelogenous leukemia (AML), which developed from severe aplastic anemia (SAA) and was successfully treated by low-dose Ara-C and aclarubicin with concomitant use of G-CSF (CAG therapy). A 37-year-old male was admitted for scrutiny of pancytopenia and diagnosed as SAA because of hypocellular bone marrow without abnormal or dysplastic cells. Although hematopoiesis recovered with steroid pulse therapy followed by administration of anabolic steroids, 29 months after initial onset of SAA, he presented as AML (FAB-M6), as his bone marrow Contained 21.6% leukemic myeloblasts and 56% of erythroblasts. Chromosome study revealed 45, XY, -7 in 14 of 20 cells analyzed. Complete remission was achieved by administration of low-dose Ara-C (20 mg/m2 for 7 days) and aclarubicin (14 mg/m2 for 4 days) along with G-CSF (200 micrograms/m2 for 7 days), without any severe complications. In the previous reports in Japan since 1982, 7 out of 8 cases with AML developing from SAA died within a year. Our results indicate that CAG therapy is useful for treatment for this subset of AML with poor prognosis.

We analyzed DNA content of resected primary colorectal carcinoma and lung metastasis by flow cytometry. Of the 14 primary lesions, 5 cases showed diploid pattern, 9 cases aneuploid pattern. In contrast, 12 metastatic lung lesions of 19 showed diploid pattern and 7 lesions aneuploid pattern. DNA index of primary and metastatic lesions was 1.4 +/- 0.4 and 1.2 +/- 0.2, respectively (p = 0.08). In combination of DNA ploidy pattern between primary and metastatic lesions, there were 11 in which ploidy pattern was identical, 1 in which metastasis was aneuploid and primary was diploid, 7 in which metastasis was diploid and primary was aneuploid. Four year, survival rate from operation of metastasis was better in diploid pattern than in aneuploid pattern, but it was not significant. These results indicate that patients who have operative indication of metastasis from colorectal carcinoma have metastatic tumor which shows relatively good biological behavior (diploid tumors) and that there are heterogeneity of ploidy pattern between primary and metastatic lesions.

We analyzed amplification, rearrangement and point mutation of the c-erbB2 oncogene about 10 cases of the stomach cancer and 30 cases of the colon cancer. Rearrangement and amplification of the c-erbB2 oncogene were found in one case of moderately differentiated tubular adenocarcinoma of the stomach. According to the restriction map, the rearrangement apparently occurred in the region from the 5' promotor to the extracellular domain. In this case, it appeared that the over expression of ErbB2 oncoprotein detected by immunohistochemical staining was associated with rearrangement of the gene. Thus it was suggested that the point mutation of the transmembrane domain, which has been demonstrated in the rat protooncogene neu, a homolog of human c-erbB2, was not detected in this case.